Characterization of Monoclonal Immunoglobulin A and G Against Shiga Toxin Binding Subunits Produced by Intranasal Immunization
Characterization of Monoclonal Immunoglobulin A and G Against Shiga Toxin Binding Subunits Produced by Intranasal Immunization
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鼻内免疫产生的针对志贺毒素结合亚基的单克隆免疫球蛋白 A 和 G 的表征
DOI：
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发表时间：
2008
影响因子：
3.7
通讯作者：
Y. Imai
中科院分区：
文献类型：
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作者：
T. Tanikawa;T. Ishikawa;T. Maekawa;K. Kuronane;Y. Imai
免疫球蛋白A （IgA）被认为在粘膜表面保护中起主要作用。然而，由于IgA类单克隆抗体（mab）的制备困难，其免疫学和生物学特性尚未得到广泛的研究。我们比较了IgA和IgG单克隆抗体抗志贺毒素B亚单位（Stx1B）的性能。这些单抗是从经重组Stx1B和霍乱毒素鼻内免疫的小鼠产生的杂交瘤中分泌出来的。ELISA结果显示，固定化Stx1B与IgA（克隆G2G7）和IgG（克隆D11C6）单抗结合的剂量响应曲线相似。通过免疫印迹分析判断，大多数IgA单抗形成二聚体，而IgG单抗为单体。IgG单抗完全抑制Stx1B与Burkitt淋巴瘤细胞株Ramos的结合，而IgA单抗仅部分抑制。IgG单抗能够中和Stx1全毒对Vero细胞的细胞毒性，而IgA单抗则不能。通过表面等离子体共振分析（包括捕获法）比较每个结合位点的结合亲和力，检测可溶性Stx1B与固定化单抗的结合。IgA单抗的关联率相似，但解离速度快了两倍，导致IgG单抗的亲和力提高了两倍。这些结果表明，人们可以获得高亲和力的IgA单抗，但毒素中和是IgA类治疗性抗体的另一个挑战。
Immunoglobulin A (IgA) is considered to play a major role in protection of the mucosal surface. However, its immunological and biological properties have not been extensively studied because the production of IgA class monoclonal antibodies (mAbs) is difficult. We compared the properties of IgA and IgG mAbs against Shiga toxin B subunits (Stx1B). These mAbs were secreted from hybridomas that had been produced from mice after intranasal immunization with recombinant Stx1B and cholera toxin. The dose response curves for the binding of the IgA (clone G2G7) and IgG (clone D11C6) mAbs to immobilized Stx1B were similar, as revealed on ELISA. The majority of the IgA mAb formed dimers while the IgG mAb was monomeric, as judged by immunoblot analysis. The IgG mAb completely inhibited the binding of Stx1B to Burkitt’s lymphoma cell line Ramos, while the inhibition by the IgA mAb was only partial. The IgG mAb was able to neutralize the cytotoxicity of Stx1 holotoxin towards Vero cells, whereas the IgA mAb was not. The binding affinity of each binding site was compared by means of surface plasmon resonance analysis involving a capture method, with which the binding of soluble Stx1B to immobilized mAb was detected. The association rate was similar but the dissociation rate was twofold faster in the case of the IgA mAb, resulting in twofold higher affinity of the IgG mAb. These results suggest that one can obtain high affinity IgA mAb but toxin neutralization is another challenge as to therapeutic antibodies of the IgA class.
影响因子：
4.4
作者：
M. Marinaro;H. Staats;T. Hiroi;R. Jackson;M. Coste;P. Boyaka;N. Okahashi;M. Yamamoto;Hiroshi Kiyono;Horst Bluethmann;K. Fujihashi;Jerry R. McGhee
通讯作者：
M. Marinaro;H. Staats;T. Hiroi;R. Jackson;M. Coste;P. Boyaka;N. Okahashi;M. Yamamoto;Hiroshi Kiyono;Horst Bluethmann;K. Fujihashi;Jerry R. McGhee